A recombinant vaccine strain of genotype vii Newcastle disease virus with hn protein mutation
A technology of Newcastle disease virus and recombinant vaccine, which is applied in the field of veterinary biological products, can solve the problems of immune failure, unable to completely prevent the spread and transmission of Newcastle disease virus, and the loss of breeding industry
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0038] Example 1: Rescue of LaSota strain
[0039] 1. Virus purification
[0040] The purification of the virus is to obtain a single virus clone. The LaSota strain was purified by the limiting dilution method. The detailed steps are as follows: Dilute the virus liquid by 10 times, and inoculate the virus liquid of each titer for 9 to 11 days after dilution SPF chicken embryos (100 μL / piece) were inoculated with 5 pieces per dilution. After 4 days, the chicken embryo allantoic fluid was harvested to measure the activity of HA. The allantoic fluid with the highest dilution factor with HA activity was selected as the next-generation purified virus fluid. The same method was used for doubling dilution and chicken embryo inoculation, and the virus was continuously purified for 5 generations, and the 5th generation virus liquid was stored in aliquots, which was used as the original seed virus for the next test.
[0041] 2. Virus sequence determination
[0042] Using the purifie...
Embodiment 2
[0061] Example 2: Rescue of recombinant Newcastle disease vaccine candidate seeds
[0062] 1. Virus screening and purification
[0063] The chicken-derived Newcastle disease virus of subtype VII.2 isolated and preserved in the laboratory was selected for reproduction and rejuvenation, and then 5 strains were purified by limited dilution method. The hemagglutination titers of different generations of each strain were See Table 5. Finally, the YN1106 / 2017 strain after 5 generations of purification was selected as the original seed virus for the construction of full-length cDNA clones.
[0064] Table 5: Hemagglutination titer table of different strains of genotype VII Newcastle disease virus at each generation
[0065]
[0066] 2. Determination and analysis of the full-length genome sequence of YN1106 / 2017 strain
[0067] Using the purified YN1106 / 2017 virus reverse transcription genome cDNA as a template, using the designed 12 pairs of primers (Table 6) to amplify the full...
Embodiment 3
[0087] Example 3 Safety test and immune effect test of rLa-VII-YN17 vaccine strain on SPF chickens
[0088] 1. Preparation of inactivated vaccine
[0089] The rLa-VII-YN17 vaccine strain was diluted 10 000 times with sterilized normal saline, inoculated with 9-11-day-old SPF chicken embryos, 0.1 mL per embryo, and incubated at 37°C. The dead embryos within 24h after inoculation were discarded, and the dead embryos were kept at 4°C in time for 24h~120h, and the mixed samples were collected. 8 , the virus titer is 10 9.65 EID 50 / 0.1mL. The Newcastle disease virus solution with the determined titer was introduced into the inactivation tank, and the final concentration of 0.1% formaldehyde solution was added to make it fully mixed, inactivated at 4°C for 48 hours, and shaken every 2 hours during the period. Oil adjuvant inactivated vaccine is prepared from the inactivated virus stock solution according to conventional methods.
[0090] 2. Safety test of overdose vaccination ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com